A reliable, reproducible flow cytometry protocol for immune cell quantification in human adipose tissue

Kerri Z Delaney; Vi Dam; Jessica Murphy; José A Morais; Ronald Denis; Henri Atlas; Radu Pescarus; Pierre Y Garneau; Sylvia Santosa

doi:10.1016/j.ab.2020.113951

A reliable, reproducible flow cytometry protocol for immune cell quantification in human adipose tissue

Anal Biochem. 2021 Jan 15:613:113951. doi: 10.1016/j.ab.2020.113951. Epub 2020 Sep 11.

Authors

Kerri Z Delaney¹, Vi Dam¹, Jessica Murphy¹, José A Morais², Ronald Denis³, Henri Atlas³, Radu Pescarus³, Pierre Y Garneau³, Sylvia Santosa⁴

Affiliations

¹ Department of Health, Kinesiology, and Applied Physiology, Concordia University, 7141 Sherbrooke St W, Montreal, QC, H4B 1R6, Canada; Metabolism, Obesity, Nutrition Lab, PERFORM Centre, Concordia University, 7200 Sherbrooke St W, Montreal, QC, H4B 1R6, Canada; Centre de Recherche - Axe Maladies Chroniques, Centre Intégré Universitaire de Santé et de Services Sociaux Du Nord-de-l'Ile-de-Montreal, Hôpital Du Sacré-Coeur de Montreal, 5400 Boul Gouin O, Montréal, QC, H4J 1C5, Canada.
² Department of Health, Kinesiology, and Applied Physiology, Concordia University, 7141 Sherbrooke St W, Montreal, QC, H4B 1R6, Canada; Division of Geriatric Medicine, McGill University Health Centre - Royal Victoria Hospital, 1001, Décarie, Montreal, Québec, H4A 3J1, Canada.
³ Departement Du Chirurgie, Hôpital Du Sacré-Coeur de Montréal, 5400 Boul Gouin O, Montréal, QC, H4J 1C5, Canada.
⁴ Department of Health, Kinesiology, and Applied Physiology, Concordia University, 7141 Sherbrooke St W, Montreal, QC, H4B 1R6, Canada; Metabolism, Obesity, Nutrition Lab, PERFORM Centre, Concordia University, 7200 Sherbrooke St W, Montreal, QC, H4B 1R6, Canada; Centre de Recherche - Axe Maladies Chroniques, Centre Intégré Universitaire de Santé et de Services Sociaux Du Nord-de-l'Ile-de-Montreal, Hôpital Du Sacré-Coeur de Montreal, 5400 Boul Gouin O, Montréal, QC, H4J 1C5, Canada. Electronic address: s.santosa@concordia.ca.

PMID: 32926866
DOI: 10.1016/j.ab.2020.113951

Abstract

The ability to accurately identify and quantify immune cell populations within adipose tissue is important in understanding the role of immune cells in metabolic disease risk. Flow cytometry is the gold standard method for immune cell quantification. However, quantification of immune cells from adipose tissue presents a number of challenges because of the complexities of working with an oily substance and the rapid deterioration of immune cell viability before analysis can be performed. Here we present a highly reproducible flow cytometry protocol for the quantification of immune cells in human adipose tissue, which overcomes these issues.

Keywords: Adipose tissue; Flow cytometry; Humans; Immune cells; Inflammation; Obesity.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adipose Tissue / immunology*
Adult
Antigens, CD / analysis
Antigens, Differentiation, Myelomonocytic / analysis
Female
Flow Cytometry / methods*
Humans
Leukocyte Common Antigens / analysis
Membrane Glycoproteins / analysis
Middle Aged
Receptors, Immunologic / analysis
Reproducibility of Results
Specimen Handling / methods

Substances

Antigens, CD
Antigens, Differentiation, Myelomonocytic
CD68 antigen, human
MRC1 protein, human
Membrane Glycoproteins
Receptors, Immunologic
Leukocyte Common Antigens
PTPRC protein, human